Background: Employees working night shifts are at a greater risk of being overweight or obese. Few studies on obesity and weight gain analyze the years of exposure to night work. The aim of this study was to determine the relationship between the years of exposure to night work and body mass index (BMI) among registered nurses. Methods: A cross-sectional analysis was performed in 18 largest public hospitals in Rio de Janeiro, Brazil. A total of 2,372 registered nurses (2,100 women) completed a comprehensive questionnaire concerning sociodemographic, professional, lifestyle, and health behavioral data. Current and past exposures to night shifts as well as BMI values were measured as continuous variables. A gamma regression model was used with an identity link function to establish the association. Results: The association between years of exposure to night work and BMI was statistically significant for both women and men after adjusting for all covariates [β = 0.036; CI95%=0.009–0.063) and β = 0.071 (CI95%=0.012–0.129), respectively]. The effect of night work was greater among men than women. For example, for those women who have worked at night for 20 years the estimated average BMI was 25.6 kg/m2 [range, 25.0–26.2]. In relation to men, after 20 years of exposure to night work the estimated average BMI was 26.9 kg/m2 [range, 25.6–28.1]. Conclusions: These findings suggest that night shift exposure is related to BMI increases. Obesity prevention strategies should incorporate improvements in work environments, such as the provision of proper meals to night workers, in addition to educational programs on the health effects of night work.